Fuyan Wang1, Weiyun Shi2, Hua Li1, Hongwei Wang3, Dapeng Sun3, Long Zhao4, Lingling Yang3, Ting Liu3, Qingjun Zhou5, Lixin Xie6. 1. Department of Ophthalmology, Clinical Medical College of Shandong University, China; State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China. 2. State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China; Eye Hospital of Shandong First Medical University, China. 3. State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China; Qingdao Eye Hospital of Shandong First Medical University, China. 4. State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China. 5. State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China; Qingdao Eye Hospital of Shandong First Medical University, China. Electronic address: qjzhou2000@hotmail.com. 6. State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China; Qingdao Eye Hospital of Shandong First Medical University, China. Electronic address: lixin_xie@hotmail.com.
Abstract
PURPOSE: Hydrogels derived from decellularized tissues provide superior biocompatibility, tenability and tissue-specific extracellular matrix (ECM) components. Based on the preparation of decellularized porcine cornea (DPC), here we developed an injectable and transparent hydrogel for the regeneration of epithelium and stroma in focal corneal defects. METHODS: The DPC-derived hydrogel was prepared with N-cyclohexyl-N'-(2-morpholinethyl) carbodiimide metho-p-toluenesulfonate/N-hydroxysuccinimide (CMC/NHS) as cross-linkers. The characteristics of the hydrogel were analyzed and its cytocompatibility was assessed by Live/Dead and Cell Counting Kit (CCK)-8 assays. Immunofluorescence staining, quantitative PCR and Western blot analyses were performed to assess the relative protein and gene expression in corneal fibroblasts on hydrogel. The safety and efficiency of the hydrogel for repairing focal corneal defects in rabbit were measured by slit-lamp, anterior segment optical coherence tomography (AS-OCT), confocal microscopy and histological analyses. RESULTS: The DPC-derived hydrogel cross-linked with CMC/NHS assumed favorable transparency, exhibited distinct mechanical properties and preserved the ECM components of native porcine cornea (NPC). In vitro experiments showed that the hydrogel maintained the phenotype, supported the proliferation and promoted the ECM synthesis of corneal fibroblasts. When injected onto rabbit corneas, the hydrogel rapidly covered, solidified and formed a smooth surface on the focal defect. Corneal epithelium was fully regenerated within 3 days. The thickness of the corneal epithelium and stroma was restored at 12 weeks after surgery without significant inflammation or scar formation. Notably, the hydrogel showed no harmful effects on the resident stroma and endothelium. CONCLUSIONS: The DPC-derived hydrogel may represent a promising biomaterial for corneal epithelial and stromal regeneration.
PURPOSE: Hydrogels derived from decellularized tissues provide superior biocompatibility, tenability and tissue-specific extracellular matrix (ECM) components. Based on the preparation of decellularized porcine cornea (DPC), here we developed an injectable and transparent hydrogel for the regeneration of epithelium and stroma in focal corneal defects. METHODS: The DPC-derived hydrogel was prepared with N-cyclohexyl-N'-(2-morpholinethyl) carbodiimidemetho-p-toluenesulfonate/N-hydroxysuccinimide (CMC/NHS) as cross-linkers. The characteristics of the hydrogel were analyzed and its cytocompatibility was assessed by Live/Dead and Cell Counting Kit (CCK)-8 assays. Immunofluorescence staining, quantitative PCR and Western blot analyses were performed to assess the relative protein and gene expression in corneal fibroblasts on hydrogel. The safety and efficiency of the hydrogel for repairing focal corneal defects in rabbit were measured by slit-lamp, anterior segment optical coherence tomography (AS-OCT), confocal microscopy and histological analyses. RESULTS: The DPC-derived hydrogel cross-linked with CMC/NHS assumed favorable transparency, exhibited distinct mechanical properties and preserved the ECM components of native porcine cornea (NPC). In vitro experiments showed that the hydrogel maintained the phenotype, supported the proliferation and promoted the ECM synthesis of corneal fibroblasts. When injected onto rabbit corneas, the hydrogel rapidly covered, solidified and formed a smooth surface on the focal defect. Corneal epithelium was fully regenerated within 3 days. The thickness of the corneal epithelium and stroma was restored at 12 weeks after surgery without significant inflammation or scar formation. Notably, the hydrogel showed no harmful effects on the resident stroma and endothelium. CONCLUSIONS: The DPC-derived hydrogel may represent a promising biomaterial for corneal epithelial and stromal regeneration.
Authors: Ghasem Yazdanpanah; Yizhou Jiang; Behnam Rabiee; Meisam Omidi; Mark I Rosenblatt; Tolou Shokuhfar; Yayue Pan; Alexandra Naba; Ali R Djalilian Journal: Tissue Eng Part C Methods Date: 2021-05 Impact factor: 3.056
Authors: Ghasem Yazdanpanah; Ritu Shah; Sri Raghurama R Somala; Khandaker N Anwar; Xiang Shen; Seungwon An; Meisam Omidi; Mark I Rosenblatt; Tolou Shokuhfar; Ali R Djalilian Journal: Ocul Surf Date: 2021-04-22 Impact factor: 6.268